This study explores the impact of multi-species co-fermentation on the thermal properties of wheat starch, emphasizing the innovative use of fish collagen as an additive. The effects of adding different levels of fish collagen (0 %, 3 %, 6 %, 9 %, 12 %, and 15 %) on the thermal properties of starch were investigated during co-fermentation with Lactobacillus plantarum and Saccharomyces cerevisiae. Utilizing analytical techniques such as X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FT-IR), we observed a significant increase in the degree of order from 1.32 to 1.55 and a notable decrease in double helix formation from 1.51 to 0.80 (p <
0.05). The introduction of collagen induced the aggregation of B-starch granules, forming aggregates approximately 16 μm in diameter. Meanwhile, collagen addition shifted the peak thermal impact by +0.91 °C and elevated the maximum mass loss rate temperature from 309.33 °C to 317.09 °C (p <
0.05). Furthermore, a decrease in peak viscosity from 4321 cp to 3123 cp and final viscosity from 4143 cp to 3171 cp (p <
0.05), along with a 3 °C increase in pasting temperature. The results suggest that fish collagen is a potential quality improver for traditional fermented starch-based foods.